This proposal concerns experiments to be conducted to analyze the genetic organization and biosynthetic characteristics of E. coli ribosomal protein formation. Studies will be conducted to examine the distribution of ribosomal proteins being synthesized in vivo and in vitro on polyribosomes after a nutritional shift up. These nascent ribosomal proteins will be fractionated and analyzed for the presence of larger precursors of the individual proteins. The translation frequency of single ribosomal proteins and the effects of different regulatory molecules on the formation of the proteins will also be examined. The characteristics of the messenger RNA molecules coding for the ribosomal proteins will be examined by isolating these RNA's from polyribosomes and purifying them by hybridization to episomal DNA molecules carrying the genetic information for some of the ribosomal proteins. We will attempt to examine the distribution, frequency of transcription, and metabolism of these ribosomal protein messenger RNA molecules. In addition, we will try to translate them into ribosomal proteins in a cell-free protein synthesizing system. We will examine the regulation of messenger RNA translation by examining the influence of ribosomal macromolecules and other cellular regulatory metabolites on the messenger activities. Finally we will attempt to isolate new conditional mutants in ribosomal proteins so that the characteristics of the genetic organization and regulation of ribosomal proteins genes can be further established.